Quantitative micro-analysis of metal ions in subcellular compartments of cultured dopaminergic cells by combination of three ion beam techniques |
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Authors: | A Carmona G Devès R Ortega |
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Institution: | (1) Groupe d’Imagerie Chimique Cellulaire et Spéciation, Chemin du solarium, Laboratoire de Chimie Nucléaire Analytique et Bioenvironnementale, Universités de Bordeaux 1 & 2, 33175 Gradignan, France;(2) Groupe d’Imagerie Chimique Cellulaire et Spéciation, Chemin du solarium, CNRS, Laboratoire de Chimie Nucléaire Analytique et Bioenvironnementale, 33175 Gradignan, France;(3) Centro Nacional de Aceleradores, Universidad de Sevilla, Parque Tecnológico Cartuja’93, 41092 Sevilla, Spain |
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Abstract: | Quantification of the trace element content of subcellular compartments is a challenging task because of the lack of analytical
quantitative techniques with adequate spatial resolution and sensitivity. Ion beam micro-analysis, using MeV protons or alpha
particles, offers a unique combination of analytical methods that can be used with micrometric resolution for the determination
of chemical element distributions. This work illustrates how the association of three ion beam analytical methods, PIXE (particle
induced X-ray emission), BS (backscattering spectrometry), and STIM (scanning transmission ion spectrometry), allows quantitative
determination of the trace element content of single cells. PIXE is used for trace element detection while BS enables beam-current
normalization, and STIM local mass determination. These methods were applied to freeze-dried cells, following a specific cryogenic
protocol for sample preparation which preserves biological structures and chemical distributions in the cells. We investigated
how iron accumulates into dopaminergic cells cultured in vitro. We found that the iron content increases in dopaminergic cells
exposed to an excess iron, with marked accumulation within distal ends, suggesting interaction between iron and dopamine within
neurotransmitter vesicles. Increased iron content of dopaminergic neurons is suspected to promote neurodegeneration in Parkinson’s
disease. |
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Keywords: | Ion beam analysis PIXE RBS Trace element Cell imaging |
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